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Comparative proteomic analysis of virulent and rifampicin attenuated Flavobacterium psychrophilum

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Flavobacterium psychrophilum is the aetiologic agent of bacterial coldwater disease and rainbow trout fry syndrome. In this study, we compared a wild-type strain (CSF 259-93) with a rifampicin-resistant strain and virulence-attenuated strain of F. psychrophilum (CSF 259-93B.17). The attenuated strain harboured a mutation in the rpoB gene consistent with resistance to rifampicin. Two-dimensional polyacrylamide gel electrophoresis (2DPAGE) and mass spectrometry demonstrated an altered proteome with eight proteins characteristic for the parent strain and six that were unique to the attenuated strain. Immunoblotting with a diagnostic monoclonal antibody (FL-43) identified a putative antigen (FP1493) that was subsequently cloned, expressed as a recombinant protein and confirmed as recognized by FL-43. 2D-PAGE, immunoblotting with rainbow trout, Oncorhynchus mykiss (Walbaum), convalescent antisera and mass spectrometry of bacterial whole-cell lysates revealed several uniquely expressed immunoreactive proteins including FP1493. An FP1493 recombinant subunit vaccine was tested, but did not provide protection against challenge with the CSF259-93 strain. While the exact mechanism responsible for altered protein synthesis and attenuation of CSF 259-93B.17 is still unknown, the differentially expressed immunoreactive proteins are a valuable resource to develop subunit vaccines and to identify proteins that are potentially involved in disease. Keywords: Attenuation, Flavobacterium psychrophilum, rifampicin, subunit vaccine.
K. Gliniewicz , K. P. Plant , S. E. LaPatra , B. R. LaFrentz , D. R. Call
Flavobacterium psychrophilum , Oncorhynchus mykiss , antigens , antiserum , disease resistance , fish diseases , gene expression , genes , immunoblotting , mass spectrometry , monoclonal antibodies , mutation , polyacrylamide gel electrophoresis , protein synthesis , proteome , proteomics , recombinant proteins , recombinant vaccines , subunit vaccines
Journal of fish diseases 2012 July v.35 no.7
Journal Articles, USDA Authors, Peer-Reviewed
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